(Cell) shRNA lentivirus production

Source

  1. /Experiment/YU Daan/shRNA/shRNA 방법.pptx
  2. /Experiment/YU Daan/shRNA/shRNA cost.xlsx

Materials

  1. pLKO.1-puro (MERCK, SHC001)
  2. AgeI (Enzynomics, R063S)
  3. EcoRI (Enzynomics, R002S)
  4. T4 DNA ligase (New England Biolabs, M0202S)
  5. MD2.G / VSV-G envelope vector (Addgene, #12259)
  6. psPAX2 / packaging vector, GAG/Pol (Addgene, #12260)
  7. Maxi Plasmid Purification kit (Thermofisher, A31217)
  8. Lipofectamine 3000 (Thermofisher, L3000008)
  9. HEK293T-Lentix cells
  10. Target cells
  11. Polybrene
  12. Selection antibiotics, for example Puromycin; concentration should be determined by target cell kill curve.

Safety

  1. Lentivirus work should be performed in BSL-2 condition using a biosafety cabinet.
  2. Collect virus-containing media, tips, tubes, and plates separately and decontaminate according to lab biosafety rules.